title: Tryptophan 121 of subunit II is the electron entry site to cytochrome-c oxidase in Paracoccus denitrificans. Involvement of a hydrophobic patch in the docking reaction.
abstract: To investigate the contribution of hydrophobic residues to the molecular recognition of cytochrome c with cytochrome oxidase, we mutated several hydrophobic amino acids exposed on subunit II of the Paracoccus denitrificans oxidase. KM and kcat values and the bimolecular rate constant were determined under steady- or presteady-state conditions, respectively. We present evidence that Trp-121 which is surrounded by a hydrophobic patch is the electron entry site to oxidase. Mutations in this cluster do not affect the binding of cytochrome c as the KM remains largely unchanged. Rather, the kcat is reduced, proposing that these hydrophobic residues are required for a fine tuning of the redox partners in the initial collisional complex to obtain a configuration optimal for electron transfer.
authors.affiliation: Molecular Genetics, Institute of Biochemistry, Marie-Curie-Strasse 9, D-60439 Frankfurt, Federal Republic of Germany.
authors.forename: H
authors.initials: H
authors.lastname: Witt
authors.forename: F
authors.initials: F
authors.lastname: Malatesta
authors.forename: F
authors.initials: F
authors.lastname: Nicoletti
authors.forename: M
authors.initials: M
authors.lastname: Brunori
authors.forename: B
authors.initials: B
authors.lastname: Ludwig
mesh_terms: D003300:Copper; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism*; D006420:Hemeproteins / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009994:Osmolar Concentration; D010231:Paracoccus denitrificans / Q000201:enzymology*; D011487:Protein Conformation; D014364:Tryptophan / Q000378:metabolism
chemical_list: D006420:Hemeproteins; D008667:Metalloproteins; D003300:Copper; D014364:Tryptophan; D003576:Electron Transport Complex IV